Lariat intronic RNAs in the cytoplasm of Xenopus tropicalis oocytes

  1. Joseph G. Gall1,2
  1. 1Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland 21218, USA
  2. 2Department of Biology, Mudd Hall, Johns Hopkins University, Baltimore, Maryland 21218, USA
  1. Corresponding author: gall{at}ciwemb.edu

Abstract

We previously demonstrated that the oocyte nucleus (germinal vesicle or GV) of Xenopus tropicalis contains a population of stable RNA molecules derived from the introns of most expressed genes. Here we show that similar stable intronic sequence (sis) RNAs occur in the oocyte cytoplasm. About 9000 cytoplasmic sisRNAs have been identified, all of which are resistant to the exonuclease RNase R. About half have been confirmed as lariat molecules and the rest are presumed to be lariats, whereas nuclear sisRNAs are a mixture of lariat and linear molecules. Cytoplasmic sisRNAs are more abundant on a molar basis than nuclear sisRNAs and are derived from short introns, mostly under 1 kb in length. Both nuclear and cytoplasmic sisRNAs are transmitted intact to the egg at GV breakdown and persist until at least the blastula stage of embryogenesis, when zygotic transcription begins. We compared cytoplasmic sisRNAs derived from orthologous genes of X. tropicalis and X. laevis, and found that the specific introns from which sisRNAs are derived are not conserved. The existence of sisRNAs in the cytoplasm of the oocyte, their transmission to the fertilized egg, and their persistence during early embryogenesis suggest that they might play a regulatory role in mRNA translation.

Keywords

Footnotes

  • Abbreviations: Dbr1, RNA lariat debranching enzyme; FPKM, fragments per kilobase per million reads; GV, germinal vesicle; Pol II, RNA polymerase II; rRNA, ribosomal RNA; RT–PCR, reverse transcription polymerase chain reaction; sis, stable intronic sequence

  • Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.045781.114.

  • Received April 10, 2014.
  • Accepted June 18, 2014.

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